This research will provide the scientific and engineering information upon which can be based the confident engineering design of automated laboratory analyses. Particular attention will be directed to the mucopolysaccharidoses-related connective tissue mucopolysaccharides (AMPs, acidic glycosoaminoglycans). The methodologies developed and the results obtained will be directly applicable to the automation of analyses for related clinical entities. Recent additions to the microscope coupled with the use of molecular probes enable biophysical cytochemical investigations on biopolymers in their native state in living cells. These physical optical methods (microspectrofluorophotometry and microspectropolarimetry) can produce physical chemical data from as little as 10 to the minus 17th power moles. These data are such as to lead directly to an engineering design for automation of cytopathology urine and body fluids analyses, and analysis of microsamples (extracts). AMPs complex with fluorescent dyes which can serve as molecular probes. The physical, chemical and optical properties of the dye-AMPs complex are characteristic of the particular complex and differ, in general, from those of the free dye and other dye-substrate complexes. The characteristic alterations in the properties of the dye underlie the rationale for this research and are the common element uniting the participating disciplines. Specifically, physical chemical and biophysical cytochemical studies using identical molecular probes will be conducted in parallel to develop rapid, accurate, qualitatve and quantitative analytical methodologies for connective tissue AMPs. Molecular probes which act as spin labels will be employed in electron spin resonance studies to gain additional insight into the environment in localized regions on these AMPs macromolecules to develop understanding of their biological behavior. Experimental systems will be set up with automation limited to essential features requiring validation. Recommendations for entering upon the engineering development and production for general use of particular automated analyses will be made as rapidly and as frequently as the research results warrant.